Investigation of Escherichia coli O157:H7 By IMS-PCR Tecniques in Poultry Slaughtering Stages

Year 2018, Volume: 13 Issue: 3, 285 - 292, 25.12.2018

Nurhan Ertaş Onmaz , Ahmet Geçer

https://doi.org/10.17094/ataunivbd.356910

Cited By: 1

Abstract

The present study was conducted to investigate the prevalence of Escherichia coli (E. coli) O157:H7 and to

determined its virulence genes (stx1 and stx2, eaeA and ehlyA) in different samples collected from a poultry cutting satages

in Samsun province, Turkey. For this purpose, 40 viscera, 40 intestinal content, 20 scalding tank water and 100 swap samples

taken from various poultry processing units (plucking machine, cutting board, removing unit of intestinal organs and

personnel hands and broiler carcass) were analysed. All samples were analyzed for the isolation of E. coli O157:H7 through

pre-enrichment in modified tryptone soy broth supplemented with novobiocin (mTSB+N) followed by immunomagnetic

separation (IMS) and selective plating on CHROM agar with Cefixim – Tellurite. Presumptive E. coli O157:H7 isolates were

tested with anti-O157 and H7 antisera. Polymerase Chain Reaction (PCR) was performed on isolates for the detection of

rfbO157, fliCH7,stx1, stx2, eaeA and ehylA genes. In this study, two intestinal content (1%) isolates were identified as E. coli

O157 while were found negative for H7. Both isolates contained stx1, stx2, eaeA and ehylA genes. As a result of this study, it

was concluded that E. coli O157 isolates detected at low levels in the broiler intestinal contents might constitute a significant

hazard for human health due to the presence of toxin genes.

Keywords

Broiler, Contamination, Escherichia coli O157:H7, Slaughterhouse

Kanatlı Kesimi Aşamalarında E. Coli O157:H7 Varlığının IMS-PZR Yöntemleri ile İncelenmesi

Abstract

Bu çalışma, Samsun ilinde kanatlı kesim aşamalarından toplanan farklı örneklerde Escherichia coli (E. coli) O157: H7

prevalansını araştırmak ve virülans genlerini (stx1 ve stx2, eaeA ve ehlyA) belirlemek amacıyla yapıldı. Bu amaçla, 40 iç organ, 40 bağırsak içeriği, 20 haşlama tank suyu ve kanatlı kesimhanesinin farklı birimlerinden (tüy yolma makinesi, kesme tahtası, iç organ çıkarma ünitesi ve personel eli ve 40 broiler karkas örneği) alınan 100 svap örneği analiz edildi. Tüm örnekler, novobiosin içeren selektif modifiye Tryptic Soy Broth (mTSB+N)’da ön zenginleştirmenin ardından immünomanyetik seperasyon (IMS) ve Cefixim – Tellurite’li CHROM agara ekilmesi yoluyla E. coli O157: H7 açısından analiz edildi. Şüpheli koloniler O157 ve H7 antiserumları ile incelendi. rfbO157, fliCH7, stx1, stx2, eaeA ve ehylA genlerinin varlığı Polimeraz Zincir Reaksiyonu (PZR) ile belirlendi. Çalışmada, 2 (%1) bağırsak içeriği örneğinden E. coli O157 identifiye edilmesine karşın, H7 tespit edilemedi. Her iki izolatın da stx1, stx2, eaeA ve ehylA geni içerdiği tespit edildi. Bu çalışma sonucunda, broiler bağırsak içeriğinde düşük oranda tespit edilen E. coli O157: H7 izolatların hepsinin anılan toksin genlerini içermesinden dolayı insan sağlığı için önemli bir tehlike oluşturabileceği kanısına varıldı.

Keywords

Broiler, Escherichia coli O157: H7, Kesimhane, Kontaminasyon

References

• 1. Adu-Gyamfi A., Torgby-Tetteh W., Appiah V., 2012. Microbiological quality of chicken sold in accra and determination of D10-value of E. coli. Food Nutr Sci, 3, 693-698. 2. Alvarez-Astorga M., Capita R., Alonso-Calleja C., Moreno B., Garcia-Fernandez MC.,2002. Microbiological quality of retail chicken by-products in Spain. Meat Sci, 62, 45-50. 3. Amara A., Badoum M., Faid M., Bouzoubaa K.,2002. Microbial contamination of poultry slaughtered in traditional shops in Morocco. MicrobiolAliments Nutr. 1994; 12: 323-327 4. Gill CO, Jones T. Effects of wearing knitted or rubber gloves on the transfer of Escherichia coli between hands and meat. J Food Prot, 65, 1045-1048. 5. Possé B., De Zutter L., Heyndrickx M., Herman L.,2007. Metabolic and genetic profiling of clinical O157 and non-O157 shiga-toxin-producing Escherichia coli. Res Microbiol, 158, 591-599. 6. Guran HS., Vural A., Erkan ME., Durmusoglu H., 2017. Prevalence and some virulence genes of Escherichia coli O157 isolated from chicken meats and giblets. Ann Anim Sci, 17(2), 555-563 7. .Abdissa R., Haile W., Fite AT., Beyi AF.,Agga GE.,Edao BM.,Tadesse F.,Korsa MG.,Beyene T., Beyene TJ.,De Zutter L.,Cox E.,Goddeeris BM.,2017. Prevalence of Escherichia coli O157:H7 in beef cattle at slaughter and beef carcasses at retail shops in Ethiopia. BMC Infect Dis, 17(1),277-283. 8. Islam MA., Mondol AS., de Boer E., Beumer RR., Zwietering MH.,Talukder KA., Heuvelink AE.,2008. Prevalence and genetic characterization of shiga toxin-producing Escherichia coli isolates from slaughtered animals in Bangladesh. Appl Environ Microbiol, 74, 5414-5421. 9. Sarimehmetoglu B., Aksoy MH., Ayaz ND., Ayaz Y., Küplülü Ö., Kaplan YZ.,2009. Detection of Escherichia coli O157:H7 in ground beef using immunomagnetic separation and multiplex PCR. Food Control, 20, 357-361. 10. Aslantaş Ö., Erdoğan S., Cantekin Z., Gülaçtı İ., Evrendilek GA.,2006. Isolation of characterization verocytotoxin-producing Escherichia coli O157 from Turkish cattle. Int J Food Microbiol,106,338-342. 11. Doyle MP., Schoeni JL.,1987. Isolation of Escherichia coli O157:H7 from retail fresh meats and poultry. Appl Environ Microbiol,53,2394-2396. 12. Hızlısoy H., Al S., Onmaz NE., Yıldırım Y., Gönülalan Z., Gümüşsoy KS., 2017. Antimicrobial resistance profiles and virulence factors of Escherichia coli O157 collected from a poultry processing plant. Turk J Vet Anim Sci, 41, 65-71. 13. Hessain AM., Al-Arfaj AA., Zakri AM., El-Jakee JK., Al-Zogibi OG., Hemeg HA., İbrahim IM., 2015. Molecular characterization of Escherichia coli O157:H7 recovered from meat and meat products relevant to human health in Riyadh, Saudi Arabia. Saudi J Biol Sci, 22(6), 725-729. 14. Heuvelink AE., Zwartkruis-Nahuis JT., Van den Biggelaar FL., Van Leeuwen WJ., De Boer E.,1999. Isolation and characterization of verocytotoxin-producing Escherichia coli O157 from slaughter pigs and poultry. Int J Food Microbiol, 52 (1-2), 67-75. 15. Kalin R., Öngör H., Çetinkaya B., 2012. Isolation and molecular characterization of Escherichia coli O157 from broiler and human samples. Foodborne Paht Dis, 9(4), 313-318. 16. Abay S., Aydin F., Ertaş N., Hizlisoy H., Erdoğdu S., Gönülalan Z., 2014. Kanatlılardan Escherichia coli O157 izolasyonu üzerine çalışmalar. Erc Univ Vet Fak Derg, 11(1), 1-6. 17. Raji MA., Minga UM., Machang’u RS.,2006. Prevalence and characterization of verotoxigenic Escherichia coli O157 isolated from local chicken in Morogoro, Tanzania. J Anim Vet Advan, 5(11), 952-958. 18. Sekmen SGD., Kaya O.,2010. Broiler piliçlerden Escherichia coli O157:H7 serotipinin identifikasyonu ve antibiyotik duyarlılıklarının belirlenmesi. Pendik Vet Mikrobiyol Derg, 37(1), 19-31. 19. Tabatabaei M., Mokarizade A., Foad-Marashi N. 2011. Detection and molecular characterization of sorbitol negative shiga toxigenic Escherichia coli in chicken from northwest of Iran. Vet Res Forum, 2(3),183-188. 20. Kagambe`ga A., Martikainen O., Lienemann T.,2012. Diarrheagenic Escherichia coli detected by 16-plex PCR in raw meat and beef intestines sold at local markets in Ouagadougou, Burkina Faso. Int J Food Microbiol, 153,154-158. 21. Beery JT., Doyle MP., Schoeni JL.,1985. Colonization of chicken caeca by Escherichia coli associated with hemorrhagic colitis. Appl Environ Microbiol, 49,310-315. 22. Baran F., Gülmez M.,2003. The occurence of Escherichia coli O157:H7 in the ground beef and chicken drum sticks. Internet J Food Safety, 2: 13-15 23. Balpetek D., Gürbüz Ü., 2010. Bazı et ürünlerinde E. coli 0157:H7 varlığının araştırılması. Eurasian J Vet Sci, 26(1), 25-31. 24. . Akkaya L., Atabay HI., Kenar B., Alisarlı M.,2006. Prevalence of verocytotoxigenic Escherichia coli O157:H7 on chicken carcasses sold in Turkey. Bull Vet Inst Pulawy, 50: 513-516. 25. Olatoye IO., Amosun EA., Ogundipe GA.,2012. Multidrug Resistant Escherichia coli O157 Contamination of Beef and Chicken in Municipal Abattoirs of Southwest Nigeria. Nat Sci, 10(8), 125-132. 26. Ojo OE., Ajuwape ATP., Otesile EB., Owoade AA., Oyekunle MA., Adetosoye AI.,2010. Potentially zoonotic shiga toxin-producing Escherichia coli serogroups in the faeces and meat of food-producing animals in Ibadan, Nigeria. Int J Food Microbiol,142,214-221. 27. Murinda SE., Nguyen LT., Landers TL., Draughon A., Mathew AG., Hogan JS., Smith LK., Hancock DD., Oliver SP.,2004. Comparison of Escherichia coli isolates from humans, food, and farm and companion animals for presence of shiga toxin-producing E. coli virulence markers. Foodborne Pathog Dis, 1, 181-184. 28. Etcheverria AI., Padola NL., Sanz ME., Polifroni R., Krüger A.,Passucci J.,Rodríguez EM.,Taraborelli AL.,Ballerio M.,Parma AE.,2010. Occurrence of shiga toxin-producing E. coli (STEC) on carcasses and retail beef cuts in the marketing chain of beef in Argentina. Meat Sci, 86, 418-421.